Bittman R, Clejan S
Department of Chemistry, Queens College of the City University of New York, Flushing 11367.
Isr J Med Sci. 1987 May;23(5):398-402.
The following modifications in the lipid composition of Mycoplasma gallisepticum or M. capricolum membranes led to changes in the rates of [14C]cholesterol and [14C]phospholipid exchange between cell membranes and an excess of small unilamellar vesicles: 1) increase in the cholesterol/phospholipid molar ratio from 0.25 to 0.92; 2) incorporation of synthetic, cross-linked phosphatidylethanolamine (PE) derivatives, 3) incorporation of sphingomyelin (SPM); and 4) increase in the phosphatidylglycerol/cardiolipin ratio of M. capricolum cell membranes by supplementing the growth medium with 0.5 mM CaCl2 CaCl2, and decrease in this ratio by supplementing the growth medium with 0.5 mM CaCl2 and 20 micrograms/ml egg phosphatidylcholine or with isopalmitic acid. The rates of radiolabeled lipid exchange decreased when the content of cholesterol, cross-linked PE, or SPM increased, indicating an inverse correlation between exchange rate and membrane lipid order. This is consistent with an exchange mechanism that involves the slow dissolution of the lipid from the surface of the donor particle. Lipidic particles appeared in trypsin-treated M. capricolum membranes obtained from cells grown with both Ca2+ and PC, whereas the hexagonal-II phase appeared in membranes from cells grown with Ca2+. Cholesterol and phospholipid exchange rates were enhanced under the conditions in which the bilayer structure was destabilized by transitional states between the lamellar and hexagonal-II phases. Thus, mycoplasmas are well suited for examination of the influence of membrane lipid composition on rates of lipid exchange between membranes.
鸡毒支原体或山羊支原体细胞膜脂质组成的以下改变导致细胞膜与过量小单层囊泡之间[14C]胆固醇和[14C]磷脂交换速率发生变化:1)胆固醇/磷脂摩尔比从0.25增加到0.92;2)掺入合成的交联磷脂酰乙醇胺(PE)衍生物;3)掺入鞘磷脂(SPM);4)通过在生长培养基中添加0.5 mM氯化钙使山羊支原体细胞膜的磷脂酰甘油/心磷脂比值增加,以及通过在生长培养基中添加0.5 mM氯化钙和20微克/毫升鸡蛋磷脂酰胆碱或异棕榈酸使该比值降低。当胆固醇、交联PE或SPM的含量增加时,放射性标记脂质的交换速率降低,表明交换速率与膜脂质有序性呈负相关。这与一种涉及脂质从供体颗粒表面缓慢溶解的交换机制一致。在用Ca2+和PC培养的细胞获得的经胰蛋白酶处理的山羊支原体细胞膜中出现脂质颗粒,而在用Ca2+培养的细胞的膜中出现六方-II相。在层状相和六方-II相之间的过渡状态使双层结构不稳定的条件下,胆固醇和磷脂的交换速率增强。因此,支原体非常适合用于研究膜脂质组成对膜间脂质交换速率的影响。